LOCUS       sequence-216967-        5452 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses N-terminus to aa168 of Flp recombinase fused to FRB CID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5452)
  AUTHORS   Weinberg BH, Cho JH, Agarwal Y, Pham NTH, Caraballo LD, Walkosz M, 
            Ortega C, Trexler M, Tague N, Law B, Benman WKJ, Letendre J, Beal J,
            Wong WW
  TITLE     High-performance chemical- and light-inducible recombinases in 
            mammalian cells and mice.
  JOURNAL   Nat Commun. 2019 Oct 24;10(1):4845. doi: 10.1038/s41467-019-12800-7.
  PUBMED    31649244
REFERENCE   2  (bases 1 to 5452)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2019-10-24"; volume: "10(1):4845. doi"; pages: " 
            10.1038/s41467-019-12800-7"
COMMENT     Sequence Label: pBW2276_pCAG-PV1-FlpO-N168-L1-FRB-NLS-BGHpA
FEATURES             Location/Qualifiers
     source          1..5452
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     466..488
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     550..569
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1223..1498
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRIS"
     CDS             1499..1519
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1529..1753
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1993..2012)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2058..2113
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2112..2131)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2474..2490)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2474..2490)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2487..2509)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2498..2514
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2522..2528,2529..2546,2547..2552))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2567..2588
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2673..2807
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2827..2961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2864..2883)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2918..2937
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3029..3046)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3200..3788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3280..3299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3959..4750,4751..4819))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4582..4601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4820..4924)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4955..5334
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
       61 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
      121 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg
      181 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
      241 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
      301 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
      361 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
      421 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
      481 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
      541 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattgatta
      601 attcgagcga aggcgcgccc attactcgca tccattctca ggctgtctcg tctcgtctcg
      661 cacgcgtgcc gccaccatga gccagttcga catcctgtgc aagacccccc ccaaggtgct
      721 ggtgcggcag ttcgtggaga gattcgagag gcccagcggc gagaagatcg ccagctgtgc
      781 cgccgagctg acctacctgt gctggatgat cacccacaac ggcaccgcca tcaagagggc
      841 caccttcatg agctacaaca ccatcatcag caacagcctg agcttcgaca tcgtgaacaa
      901 gagcctgcag ttcaagtaca agacccagaa ggccaccatc ctggaggcca gcctgaagaa
      961 gctgatcccc gcctgggagt tcaccatcat cccttacaac ggccagaagc accagagcga
     1021 catcaccgac atcgtgtcca gcctgcagct gcagttcgag agcagcgagg aggccgacaa
     1081 gggcaacagc cacagcaaga agatgctgaa ggccctgctg tccgagggcg agagcatctg
     1141 ggagatcacc gagaagatcc tgaacagctt cgagtacacc tccggagggt ctggctccgg
     1201 atcaagtggt ggcagcggta ccatcctctg gcatgagatg tggcatgaag gcctggaaga
     1261 ggcatctcgt ttgtactttg gggaaaggaa cgtgaaaggc atgtttgagg tgctggagcc
     1321 cttgcatgct atgatggaac ggggccccca gactctgaag gaaacatcct ttaatcaggc
     1381 ctatggtcga gatttaatgg aggcccaaga gtggtgcagg aagtacatga aatcagggaa
     1441 tgtcaaggac ctcctccaag cctgggacct ctattatcat gtgttccgac gaatctcacc
     1501 caagaaaaag cggaaggtgt gagaattcct gtgccttcta gttgccagcc atctgttgtt
     1561 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1621 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1681 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     1741 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     1801 gcgccctgta gcgtgtaccg ctgggagttc gtagacggaa acaaacgcag aatccaagcg
     1861 cggccgccca ggatacatag attaccacaa ctccgagccc ttccaccgct agcgcactcc
     1921 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     1981 ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     2041 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     2101 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     2161 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg
     2221 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     2281 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     2341 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     2401 actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct
     2461 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2521 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2581 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2641 ggatccgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccagttccgc
     2701 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     2761 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     2821 aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2881 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2941 gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3001 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3061 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3601 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3841 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3901 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4081 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4381 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4561 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4621 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4801 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4861 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4921 cgcgcacatt tccccgaaaa gtgccacctg ggtcgacatt gattattgac tagttattaa
     4981 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     5041 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     5101 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     5161 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     5221 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     5281 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag
     5341 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
     5401 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg gg
//
